Copper binding dynamics and olfactory impairment in fathead minnows (Pimephales promelas).

When fish are exposed to sublethal, environmentally relevant Cu concentrations, olfactory acuity is impaired. The goals of the present study were to investigate the binding dynamics of waterborne Cu in the olfactory epithelium (OE), to examine the influence of calcium (Ca(2+)) on Cu binding, and to link Cu-OE binding to changes in olfactory acuity. Using short-term in vivo waterborne exposures to (64)Cu, we found that Cu accumulates rapidly in the OE, reaching a plateau by 3 h. The binding affinity (log K(Cu-OE)) and binding capacity (B(max)) of (64)Cu in the OE were 6.7 and 10.0 nmol Cu g(-1), respectively. As waterborne Ca(2+) was increased from 50 to 1000 microM L(-1), the B(max) of Cu decreased by approximately 50% while the log K(Cu-OE) remained constant, indicative of noncompetitive inhibition. Using electro-olfactograms (EOG), short-term exposures to 160 and 240 nmol Cu L(-1) were found to reduce olfactory responses to 10(-5) M l-arginine by 72 and 79%, respectively. Short-term exposure to 160 nmol Cu L(-1) also caused a 15-fold reduction in behavioral responses to a food stimulus. Interestingly, increasing waterborne Ca(2+) did not reduce the effects of Cu on EOG or behavioral responses. These results demonstrate that short-term, environmentally realistic concentrations of Cu not only bind to the OE of fathead minnows but also impair their olfactory sensitivity and behavioral responses to olfactory stimuli. Waterborne Ca(2+) reduces Cu-OE binding but does not protect against olfactory impairment.

Kin recognition and cannibalistic behaviours by adult male fathead minnows (Pimephales promelas).

Parental care is an energetically demanding activity that ensures genes are efficiently passed from one generation to the next. According to evolutionary theory, the greatest energetic investment should be directed towards offspring that are most closely related to the parent. Male fathead minnows, Pimephales promelas, provide this parental investment to developing embryos but not newly hatched larvae. Therefore, selection should favour recognition of embryonic kin to ensure energetic expenditure is optimally invested. In this study, adult male fathead minnows were tested using behavioural assays, with egg cannibalism as an endpoint, to determine whether adult males could discriminate between related and unrelated embryos. Egg cannibalism was highest when adult male fathead minnows were presented with unrelated eggs and lowest when presented with eggs fertilized by the test subject (related eggs). The degree of cannibalism was also a function of breeding status. Unrelated males in breeding condition showed an intermediate response between the low cannibalism demonstrated by related males and the high cannibalism demonstrated by unrelated males in a nonbreeding condition. These results suggest that although male fathead minnows can discriminate between unrelated and related embryos, at least some component of parental investment is a simple function of breeding status.

Epidermal 'alarm substance' cells of fishes maintained by non-alarm functions: possible defence against pathogens, parasites and UVB radiation.

Many fishes possess specialized epidermal cells that are ruptured by the teeth of predators, thus reliably indicating the presence of an actively foraging predator. Understanding the evolution of these cells has intrigued evolutionary ecologists because the release of these alarm chemicals is not voluntary. Here, we show that predation pressure does not influence alarm cell production in fishes. Alarm cell production is stimulated by exposure to skin-penetrating pathogens (water moulds: Saprolegnia ferax and Saprolegnia parasitica), skin-penetrating parasites (larval trematodes: Teleorchis sp. and Uvulifer sp.) and correlated with exposure to UV radiation. Suppression of the immune system with environmentally relevant levels of Cd inhibits alarm cell production of fishes challenged with Saprolegnia. These data are the first evidence that alarm substance cells have an immune function against ubiquitous environmental challenges to epidermal integrity. Our results indicate that these specialized cells arose and are maintained by natural selection owing to selfish benefits unrelated to predator-prey interactions. Cell contents released when these cells are damaged in predator attacks have secondarily acquired an ecological role as alarm cues because selection favours receivers to detect and respond adaptively to public information about predation.

Brook char (Salvelinus fontinalis) can differentiate chemical alarm cues produced by different age/size classes of conspecifics.

A wide diversity of aquatic organisms release chemical alarm cues when captured by a predator. For most animals, it is not known whether the specific chemicals that comprise the alarm cue are conserved as prey animals age. In this study, we tested whether brook char (Salvelinus fontinalis) can differentiate alarm cues produced by individuals of different ages/sizes. In separate laboratory experiments we exposed small brook char and large brook char to chemical alarm cues from small brook char, large brook char, and a control of swordtails (Xiphophorus helleri). Both small and large brook char responded with antipredator behavior to chemical alarm cues from both small and large char, but not to those from swordtails. Small char responded with a greater response intensity to cues of small char than to cues of large char. In contrast, large char responded with a greater response intensity to cues of large char than to cues of small char. These results suggest that chemical(s) that act as the alarm cue for fish of different age/size classes may be: (1) identical and that there may be other chemical(s) that allow the test fish to distinguish between cues from fish of different ages/sizes, or (2) the cues are not identical, but similar enough to be recognized.